Lipid-free NisI: interaction with nisin and contribution to nisin immunity via secretion

Takala, Timo M.; Koponen, Olli; Qiao, Mingqiang; Saris, Per E. J.

doi:10.1016/j.femsle.2004.06.032

Cited by 14 publications

(27 citation statements)

References 25 publications

(32 reference statements)

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“…Nisin Binding-Previously it was shown by native PAGE that NisI directly binds to nisin (24) although the interaction is rather weak with a K D for the interaction in the micromolar range (25). Stein et al (24) also showed that NisI does not bind the structurally closely related subtilin.…”

Section: Resultsmentioning

confidence: 99%

“…It was also shown by native PAGE that in vitro NisI specifically binds to nisin but not subtilin (24). A nisin binding affinity in the low micromolar range was determined for lipid-free NisI by surface plasmon resonance (25). Furthermore, experiments with C-terminal truncation variants of NisI suggested that the C-terminal 22 amino acids of NisI are involved in nisin immunity (26,27).…”

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confidence: 99%

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The Solution Structure of the Lantibiotic Immunity Protein NisI and Its Interactions with Nisin

Hacker

Christ

Duchardt‐Ferner

et al. 2015

Journal of Biological Chemistry

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Background:The lipoprotein NisI confers immunity to Lactococcus lactis against its own lantibiotic nisin. Results: NisI contains a membrane and a nisin binding domain both with the same fold as SpaI from Bacillus subtilis. Conclusion: The C-terminal domain of NisI interacts directly and specifically with nisin. Significance: Our results help to understand the mechanism of the NisI mediated immunity against nisin on a structural level.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

The Solution Structure of the Lantibiotic Immunity Protein NisI and Its Interactions with Nisin

Hacker

Christ

Duchardt‐Ferner

et al. 2015

Journal of Biological Chemistry

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“…The equilibrium dissociation constant (1.2 M) for the binding interaction between HalI and HalC8 evaluated by the SPR assay was similar to that for the NisInisin interaction (range, 0.6 to 2 M) (30). The moderately strong binding may be helpful for releasing the captured peptide antibiotic into the environment again by some unknown mechanism.…”

Section: Discussionmentioning

confidence: 56%

The Helix-Loop-Helix Motif at the N Terminus of HalI Is Essential for Its Immunity Function against Halocin C8

et al. 2008

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Halocin C8 (HalC8) is a stable microhalocin exhibiting strong antimicrobial activity against a wide range of haloarchaea. HalI, a 207-amino-acid peptide derived from the N terminus of the HalC8 preproprotein, is the immunity protein of HalC8. In this study, the molecular mechanism of the immunity function of HalI was investigated. Both pull-down and surface plasmon resonance assays revealed that HalI directly interacted with HalC8, and a mixture of purified HalI and HalC8 readily formed a heterocomplex, which was verified by gel filtration. Interestingly, HalC8 tended to form a self-associated complex, and one immunity protein likely sequestered multiple halocins. Significantly, the helix-loop-helix (HLH) motif containing a 4-amino-acid repeat (RELA) at the N terminus of HalI played a key role in its immunity activity. Disruption of the HLH motif or mutagenesis of the key residues of the RELA repeat resulted in loss of both the immunity function and the ability of HalI to bind to HalC8. These results demonstrated that HalI sequestered the activity of HalC8 through specific and direct binding.Halocins are proteinaceous antibiotics that are ribosomally synthesized by extremely halophilic archaea and excreted into the environment to kill or inhibit other haloarchaeal cells (15). Halocins were first discovered in 1982 (23) and later were found to be produced by most rod-shaped haloarchaea (12, 32). So far, several halocins have been purified and characterized (7,11,15,17,31), and three halocin genes coding for H4 (halH4), S8 (halS8), and C8 (proC8) have been cloned (2,19,29). Although the amino acid sequences of these halocins are quite diverse, the halocins share many features. For example, halocins are usually produced upon transition from the exponential to stationary phase and are processed from larger precursor proteins (2,19,29). In addition, halocins appear to be exported by a twin-arginine translocation (Tat) pathway (24, 29), as their precursors usually contain a Tat signal at their N termini (2,19,29). Accordingly, halocin has been considered a good model for examining gene expression regulation and protein processing and transportation in haloarchaea.HalC8 is an extremely stable microhalocin with a wide inhibition spectrum (7). The HalC8 gene encodes a 283-aminoacid preproprotein (ProC8), which is processed into two functional peptides, the C-terminal peptide antibiotic HalC8 (76 amino acids) and the N-terminal immunity protein HalI (29). As far as we know, HalI is the only halocin immunity protein that has been characterized. It is likely oriented toward the outside of the cellular membrane and hence inhibits HalC8 by sequestration (29). However, the molecular details of the interaction between HalI and HalC8 have not been elucidated yet. In this study, we demonstrated that HalI blocked HalC8 activity by direct interaction with HalC8. The motif and residues of HalI required for this functional interaction were also determined. MATERIALS AND METHODSStrains and growth conditions. Escherichia coli DH5␣ (Clo...

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“…When lantibiotics are considered, nisin A is the only one that has been subjected to a considerable amount of research concerning resistance (13,14,15,22,23,(29)(30)(31). Very little is known about resistance to other lantibiotics, such as gallidermin, although work has been done to show how resistance works in the producer strain of the antibiotic (17).…”

Section: Discussionmentioning

confidence: 99%

Oxidation of Lanthionines Renders the Lantibiotic Nisin Inactive

Wilson-Stanford

Kalli

Håkansson

et al. 2009

Appl Environ Microbiol

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The peptide antibiotic nisin A belongs to the group of antibiotics called lantibiotics. They are classified as lantibiotics because they contain the structural group lanthionine. Lanthionines are composed of a single sulfur atom that is linked to the ␤-carbons of two alanine moieties. These sulfur atoms are vulnerable to environmental oxidation. A mild oxidation reaction was performed on nisin A to determine the relative effects it would have on bioactivity. High-mass-accuracy Fourier transform ion cyclotron resonance mass spectrometry data revealed the addition of seven, eight, and nine oxygens. These additions correspond to the five lanthionines, two methionines, and two histidines that would be susceptible to oxidation. Subsequent bioassays revealed that the oxidized form of nisin A had a complete loss of bactericidal activity. In a competition study, the oxidized nisin did not appear to have an antagonistic affect on the bioactivity of nisin A, since the addition of an equal molar concentration of the oxidized variant did not have an influence on the bactericidal activity of the native antibiotic. Electron microscopy data revealed that the oxidized forms were still capable of assembling into large circular complexes, demonstrating that oxidation does not disrupt the lateral assembly mechanism of the antibiotic. Affinity thin-layer chromatography and fluorescence microscopy experiments suggested that the loss of activity is due to the inability of the oxidized form of nisin to bind to the cell wall precursor lipid II. Given the loss of bioactivity following oxidation, oxidation should be an important factor to consider in future production, purification, pharmacokinetic, and pharmacodynamic studies.

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Lipid-free NisI: interaction with nisin and contribution to nisin immunity via secretion

Cited by 14 publications

References 25 publications

The Solution Structure of the Lantibiotic Immunity Protein NisI and Its Interactions with Nisin

The Solution Structure of the Lantibiotic Immunity Protein NisI and Its Interactions with Nisin

The Helix-Loop-Helix Motif at the N Terminus of HalI Is Essential for Its Immunity Function against Halocin C8

Oxidation of Lanthionines Renders the Lantibiotic Nisin Inactive

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